LOCUS       sequence-431505-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Pol 2 promoter) ubiquitous expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 5126)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA368
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        444..2956
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     primer_bind     complement(3036..3052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3036..3052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3049..3071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3060..3076
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3084..3090,3091..3108,3109..3114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3129..3150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3267..3284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3438..4026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3518..3537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4204..5013)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4927..4946
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5005..5024
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacgta caccatgaat ggcatcaact ctgaatcaaa tctttgcaga
      481 tgcacctact tctcatttcc actgtcacat catttttcca gatctcgctg cctgttatgt
      541 ggcccacaaa ccaagacacg ttttatggcc attaaagctg gctgatcgtc gccaaacacc
      601 aaatacatat caatatgtac attcgagaaa gaagcgatca aagaagcctc ttcgggcgag
      661 taggagaatg cggaggagaa ggagaacgag ctgatctagt atctctccac aatccaatgc
      721 caactgacca actggccata ttcggagcaa tttgaagcca atttccatcg cctggcgatc
      781 gctccattct tggctatatg tttttcaccg ttcccggggc cattttcaaa gactcgtcgg
      841 taagataaga ttgtgtcact cgctgtctct cttcatttgt cgaagaatgc tgaggaattt
      901 cgcgatgacg tcggcgagta ttttgaagaa tgagaataat ttgtatttat acgaaaatca
      961 gttagtggaa ttttctacaa aaacatgtta tctatagata attttgttgc aaaatatgtt
     1021 gactatgaca aagattgtat gtatatacct ttaatgtatt ctcattttct tatgtattta
     1081 taatggcaat gatgatactg atgatatttt aagatgatgc cagaccacag gctgatttct
     1141 gcgtcttttg ccgaacgcag tgcatgtgcg gttgttgttt tttggaatag tttcaatttt
     1201 cggactgtcc gctttgattt cagtttcttg gcttattcaa aaagcaaagt aaagccaaaa
     1261 aagcgagatg gcaataccaa atgcggcaaa acggtagtgg aaggaaaggg gtgcggggca
     1321 gcggaaggaa gggtggggcg gggcgtggcg gggtctgtgg ctgggcgcga cgtcaccgac
     1381 gttggagcca ctcctttgac catgtgtgcg tgtgtgtatt attcgtgtct cgccactcgc
     1441 cggttgtttt tttcttttta tctcgctctc tctagcgcca tctcgtacgc atgctcaacg
     1501 caccgcatgt tgccgtgtcc tttatgcgtc attttggctc gaaataggca attatttaaa
     1561 caaagattag tcaacgaaaa cgctaaaata aataagtcta caatatggtt acttattgcc
     1621 atgtgtgtgc agccaacgat agcaacaaaa gcaacaacac agtggctttc cctctttcac
     1681 tttttgtttg caagcgcgtg cgagcaagac ggcacgaccg gcaaacgcaa ttacgctgac
     1741 aaagagcaga cgaagttttg gccgaaaaac atcaaggcgc ctgatacgaa tgcatttgca
     1801 ataacaattg cgatatttaa tattgtttat gaagctgttt gacttcaaaa cacacaaaaa
     1861 aaaaaataaa acaaattatt tgaaagagaa ttaggaatcg gacagcttat cgttacgggc
     1921 taacagcaca cccagacgaa atagcttacc tgacgtcaca gcctctggaa gaactgccgc
     1981 caagcagaga gagagagaaa aagagggaga gcagcttaga ccgcatgtgc ttgtgtgtga
     2041 ggggtctctc tcttcgtctg ctgtttgcgc aaacgcatag actgcactga gaaaatcgat
     2101 tacctatttt ttatgaatga atatttgcac tattactatt caaaactatt aagatagcaa
     2161 tcacattcaa tagccaaata ctataccacc tgagcgatgc aacgaaatga tcaatttgag
     2221 caaaaatgct gcatatttag gacggcatca ttatagaaat gcttcttgct gtgtactttt
     2281 ctctcgtctg gcagctgttt cgccgttatt gttaaaaccg gcttaagtta ggtgtgtttt
     2341 ctacgactag tgatgcccct actagaagat gtgtgttgca caaatgtccc tgaataacca
     2401 atttgaagtg cagatagcag taaacgtaag ctaatatgaa tattatttaa ctgtaatgtt
     2461 ttaatatcgc tggacattac taataaaccc actataaaca catgtacata tgtatgtttt
     2521 ggcatacaat gagtagttgg ggaaaaaatg tgtaaaagca ccgtgaccat cacagcataa
     2581 agataaccag ctgaagtatc gaatatgagt aacccccaaa ttgaatcaca tgccgcaact
     2641 gataggaccc atggaagtac actcttcatg gcgatataca agacacacac aagcacgaac
     2701 acccagttgc ggaggaaatt ctccgtaaat gaaaacccaa tcggcgaaca attcataccc
     2761 atatatggta aaagttttga acgcgacttg agagcggaga gcattgcggc tgataaggtt
     2821 ttagcgctaa gcgggcttta taaaacgggc tgcgggacca gttttcatat cactaccgtt
     2881 tgagttcttg tgctgtgtgg atactcctcc cgacacaaag ccgctccatc agccagcagt
     2941 cgtctaatcc agagaccccg gatcggagga gcgtcttcat cggatcccgg gcccgtcgac
     3001 tgcagaggcc tgcatgcaag cttggtgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4141 taaaaatgaa gttttaaatc aagcccaatc tgaataatgt tacaaccaat taaccaattc
     4201 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     4261 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     4321 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     4381 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     4441 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     4501 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     4561 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     4621 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     4681 atattcttct aatacctgga atgctgtttt tccggggatc gcagtggtga gtaaccatgc
     4741 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     4801 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     4861 aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc
     4921 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     4981 cggcctcgac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     5041 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     5101 attttgagac acgggccaga gctgca
//
